Characterization of a secreted cystatin of the parasitic nematode Haemonchus contortus and its immune-modulatory effect on goat monocytes

Background: haemonchosis is a disease of the small ruminant caused by a nematode parasite haemonchus contortus,and it is most important and alarming challenges to the small ruminant's production. the infection of the h. contortus could cause high economic losses worldwide. h. contortus is a blood feeding parasite which penetrates into the abomasal mucosa to feed the blood of the host and causing the anemia and decreased total plasma protein. modulation and suppression of the immune response of the host by nematode parasites have been reported extensively,and the cysteine protease inhibitor (cystatin) is identified as one of the major immunomodulators. methods: the recombinant protein of hccyst-3 was expressed in a histidine-tagged fusion soluble form in escherichia coli,and its inhibitory activity against cathepsin l,b,as well as papain,were identified by fluorogenic substrate analysis. native hccyst-3 protein was localized by an immunohistochemical test. the immunomodulatory effects of hccyst-3 on cytokine secretion,mhc molecule expression,no production and phagocytosis were observed by co-incubation of rhccyst-3 with goat monocytes. results: we cloned and produced recombinant cystatin protein from h. contortus (rhccyst-3) and investigated its immunomodulatory effects on goat monocyte. the rhccyst-3 protein is biologically functional as shown by its ability to inhibit the protease activity of cathepsin l,cathepsin b,and papain. the immunohistochemical test demonstrated that the native hccyst-3 protein was predominantly localized at the body surface and internal surface of the worm's gut. we demonstrated that rhccyst-3 could be distinguished by antisera from goat experimentally infected with h. contortus and could uptake by goat monocytes. the results showed that the engagement of rhccyst-3 decreased the production of tnf-α,il-1β and il-12p40. however,it significantly increased the secretion of il-10 and tgf-β1 in goat monocytes. after rhccyst-3 exposure,the expression of mhc-ii on goat monocytes was restricted. moreover,rhccyst-3 could upregulate lps induced no production of goat monocytes. phagocytotic assay by fitc-dextran internalization showed that rhccyst-3 inhibited the phagocytosis of goat monocytes. conclusions: our results suggested that the recombinant cystatin from h. contortus (rhccyst-3) significantly modulated goat monocyte function in multiple aspects. © 2017 the author(s).